Browse > Article
http://dx.doi.org/10.7314/APJCP.2012.13.9.4807

Characterization and Resistance Mechanisms of A 5-fluorouracil-resistant Hepatocellular Carcinoma Cell Line  

Gu, Wei (Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University)
Fang, Fan-Fu (Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University)
Li, Bai (Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University)
Cheng, Bin-Bin (Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University)
Ling, Chang-Quan (Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.13, no.9, 2012 , pp. 4807-4814 More about this Journal
Abstract
Purpose: The chemoresistance of human hepatocellular carcinoma (HCC) to cytotoxic drugs, especially intrinsic or acquired multidrug resistance (MDR), still remains a major challenge in the management of HCC. In the present study, possible mechanisms involved in MDR of HCC were identified using a 5-fluorouracil (5-FU)-resistant human HCC cell line. Methods: BEL-7402/5-FU cells were established through continuous culturing parental BEL-7402 cells, imitating the pattern of chemotherapy clinically. Growth curves and chemosensitivity to cytotoxic drugs were determined by MTT assay. Doubling times, colony formation and adherence rates were calculated after cell counting. Morphological alteration, karyotype morphology, and untrastructure were assessed under optical and electron microscopes. The distribution in the cell cycle and drug efflux pump activity were measured by flow cytometry. Furthermore, expression of potential genes involved in MDR of BEL-7402/5-FU cells were detected by immunocytochemistry. Results: Compared to its parental cells, BEL-7402/5-FU cells had a prolonged doubling time, a lower mitotic index, colony efficiency and adhesive ability, and a decreased drug efflux pump activity. The resistant cells tended to grow in clusters and apparent changes of ultrastructures occurred. BEL-7402/5-FU cells presented with an increased proportion in S and G2/M phases with a concomitant decrease in G0/G1 phase. The MDR phenotype of BEL-7402/5-FU might be partly attributed to increased drug efflux pump activity via multidrug resistance protein 1 (MRP1), overexpression of thymidylate synthase (TS), resistance to apoptosis by augmentation of the Bcl-xl/Bax ratio, and intracellular adhesion medicated by E-cadherin (E-cad). P-glycoprotein (P-gp) might play a limited role in the MDR of BEL-7402/5-FU. Conclusion: Increased activity or expression of MRP1, Bcl-xl, TS, and E-cad appear to be involved in the MDR mechanism of BEL-7402/5-FU.
Keywords
Hepatocellular carcinoma; multidrug resistance; 5-fluorouracil; ATP-binding cassette transporters;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Baguley BC (2010). Multidrug resistance in cancer. Methods Mol Biol, 596, 1-14.   DOI
2 Bruix J, Sherman M; Practice Guidelines Committee, American Association for the Study of Liver Diseases (2005). Management of hepatocellular carcinoma. Hepatology, 42, 1208-36.   DOI   ScienceOn
3 Chen R, Zhu D, Ye X, et al (1980). Establishment of three human liver carcinoma cell lines and some of their biological characteristics in vitro. Sci Sin, 23, 236-47.
4 Dai ZQ, Wang R, Ling SK, et al (2007). Simulated microgravity inhibits the proliferation and osteogenesis of rat bone marrow mesenchymal stem cells. Cell Prolif, 40, 671-84.   DOI   ScienceOn
5 Durand RE, Olive PL (1981). Flow cytometry studies of intracellular adriamycin in single cells in vitro. Cancer Res, 41(9 Pt 1), 3489-94.
6 El-Serag HB (2011). Hepatocellular carcinoma. N Engl J Med, 365, 1118-27.   DOI   ScienceOn
7 Ferlay J, Shin HR, Bray F, et al (2010). GLOBOCAN 2008, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10 [Internet]. Lyon, France: International Agency for Research on Cancer. Available from: http://globocan.iarc.fr
8 Frustaci S, Bearz A, Basso B, et al (2003). Efficacy of oxaliplatin and 5-fluorouracil in hepatocarcinoma (HCC). Proc Am Soc Clin Oncol, 22, (abstr 1346).
9 Green SK, Francia G, Isidoro C, Kerbel RS (2004). Antiadhesive antibodies targeting E-cadherin sensitize multicellular tumor spheroids to chemotherapy in vitro. Mol Cancer Ther, 3, 149-59.
10 Guglielmi A, Aschele C, Grossi F, et al (1996). Alternating bolus and continuous infusion 5-fluorouracil: a strategy to overcome resistance to this fluoropyrimidine in advanced colorectal cancer patients. Cytotechnology, 19, 215-9.   DOI
11 Han KQ, Huang G, Gu W, et al (2007). Anti-tumor activities and apoptosis-regulated mechanisms of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice. World J Gastroenterol, 13, 3374-9.
12 Huai-Yun H, Secrest DT, Mark KS, et al (1998). Expression of multidrug resistance-associated protein (MRP) in brain microvessel endothelial cells. Biochem Biophys Res Commun, 243, 816-20.   DOI   ScienceOn
13 Jianmin Z, Hongfang W, Meifu F (2002). Resistance of multicellular aggregates to pharmorubicin observed in human hepatocarcinoma cells. Braz J Med Biol Res, 35, 255-60.   DOI
14 Jin J, Huang M, Wei HL, Liu GT (2002). Mechanism of 5-fluorouracil required resistance in human hepatocellular carcinoma cell line Bel(7402). World J Gastroenterol, 8, 1029-34.
15 Li B, Ye T, Zhao L, et al (2006). Effects of multidrug resistance, antisense RNA on the chemosensitivity of hepatocellular carcinoma cells. Hepatobiliary Pancreat Dis Int, 5, 552-9.
16 Marin JJ, Romero MR, Briz O (2010). Molecular bases of liver cancer refractoriness to pharmacological treatment. Curr Med Chem, 17, 709-40.   DOI
17 Li G, Chen X, Wang Q, et al (2007). The roles of four multi-drug resistance proteins in hepatocellular carcinoma multidrug resistance. J Huazhong Univ Sci Technolog Med Sci, 27, 173-5.   DOI
18 Li YX, Lin ZB, Tan HR (2004). Wild type p53 increased chemosensitivity of drug-resistant human hepatocellular carcinoma Bel7402/5-FU cells. Acta Pharmacol Sin, 25, 76-82.
19 Livraghi T, Makisalo H, Line PD (2011). Treatment options in hepatocellular carcinoma today. Scand J Surg, 100, 22-9.
20 Meta-analysis Group In Cancer (1998). Efficacy of intravenous continuous infusion of fluorouracil compared with bolus administration in advanced colorectal cancer. J Clin Oncol, 16, 301-8.
21 Petriz J, Garcia-Lopez J (1997). Flow cytometric analysis of P-glycoprotein function using rhodamine 123. Leukemia, 11, 1124-30.   DOI
22 Qin S, Thongprasert S, Lim HY, et al (2010). Oxaliplatin + 5-fluorouracil/leucovorin (FOLFOX4) in advanced HCC: updated efficacy results of the EACH trial. Ann Oncol, 21(Suppl 8), 826P.   DOI
23 Ryu SH, Chung YH (2010). How to overcome multidrug resistance in chemotherapy for advanced hepatocellular carcinoma. Liver Int, 30, 496-8.   DOI
24 Shaffer LG, Tommerup N, ed (2005). ISCN 2005: An International System for Human Cytogenetic Nomenclature. S. Karger, Basel, Switzerland.
25 Sun Z, Zhao Z, Li G, et al (2010). Relevance of two genes in the multidrug resistance of hepatocellular carcinoma: in vivo and clinical studies. Tumori, 96, 90-6.
26 Wrzesinski SH, Taddei TH, Strazzabosco M (2011). Systemic therapy in hepatocellular carcinoma. Clin Liver Dis, 15, 423-41.   DOI
27 Tong SW, Yang YX, Hu HD, et al (2012). Proteomic investigation of 5-fluorouracil resistance in a human hepatocellular carcinoma cell line. J Cell Biochem, 113, 1671-80.
28 Uchibori K, Kasamatsu A, Sunaga M, et al (2012). Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines. Int J Oncol, 40, 1005-10.
29 Wilson TR, Johnston PG, Longley DB (2009). Anti-apoptotic mechanisms of drug resistance in cancer. Curr Cancer Drug Targets, 9, 307-19.   DOI
30 Yan F, Wang XM, Liu ZC, et al (2010). JNK1, JNK2, and JNK3 are involved in P-glycoprotein-mediated multidrug resistance of hepatocellular carcinoma cells. Hepatobiliary Pancreat Dis Int, 9, 287-95.
31 Yang JX, Luo Y, Qiu HM, Tang WX (2009). Characterization and resistance mechanisms of cisplatin-resistant human hepatocellular carcinoma cell line. Saudi Med J, 30, 35-40.
32 Yunis JJ (1981). New chromosome techniques in the study of human neoplasia. Hum Pathol, 12, 540-9.   DOI
33 Zhai BJ, Shao ZY, Zhao CL, et al (2006). Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice. World J Gastroenterol, 12, 6614-9.
34 Zhou J, Cheng SC, Luo D, Xie Y (2001). Study of multi-drug resistant mechanisms in a taxol-resistant hepatocellular carcinoma QGY-TR 50 cell line. Biochem Biophys Res Commun, 280, 1237-42.   DOI
35 Zhou Y, Ling XL, Li SW, et al (2010). Establishment of a human hepatoma multidrug resistant cell line in vitro. World J Gastroenterol, 16, 2291-7.   DOI